EP0077109A2
Expired / Lapsed
EP
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DNA molecules comprising the genes for preprochymosin and its maturation forms, and microorganisms transformed thereby
Assignee: Unilever PLC
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Filing: 1982-10-13
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Granted: —
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Expires: 2002-10-13
Abstract
Abstract The present invention relates to recombinant DNA and plasmids comprising specific structural genes of mammalian origin coding for the various allelic and maturation forms of preprochymosin, particularly those of bovine origin, and the use of said recombinant plasmids to transform microorganisms in which said genes are expressed.
Inventors
- 👤 Jan Maat
- 👤 Cornelis Theodorus Verrips
- 👤 Adrianus Marinus Ledeboer
- 👤 Luppo Edens
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 17:30
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6478; C12N9/6481; C12N15/00; C12N15/09; C12N15/10; C12N15/102; C12N15/63; C12N15/70; C12N15/71; C12N9/52; C12N9/58; C12N9/60
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
111 words
A process for producing DNA sequences encoding various allelic c and maturation forms of preprochymosin according to Fig. 1 and 2 or Fig. 19-22, characterized in that : (a) the message RNA (mRNA) of preprochymosin is isolated and purified; (b) mRNA is converted into double stranded DNA (dsDNA); (c) said dsDNA is incorporated in an appropriate plasmid; (d) said plasmid is transferred into bacterial cells and hybrid plasmid DNA molecules are selected to yield a plasmid containing a substantially complete copy of the preprochymosin mRNA; (e) this DNA is further genetically engineered so as to code exactly for preprochymosin or its maturation forms and brought under transcriptional control of appropriate regulons.
Claim 2
Dependent Claim
55 words
A process for producing a recombinant plasmid, characterized by combining (i) a structural gene coding for the various allelic and maturation forms of mammalian preprochymosin according to Fig. 1 and 2 or 19-22; with (ii) specific DNA sequences regulating the expression of said gene in a microbial host, consisting of an inducible or constitutive regulon.
Claim 3
Dependent Claim
22 words
A process according to Claim 2, characterized in that the regulon consists of a double lac UV5 system according to Fig. 8.
Claim 4
Dependent Claim
43 words
A process according to Claim 2, characterized in that a regulon is used which consists of at least one modified tryptophan system according to Fig. 9, wherein the information coding for the trp attenuator protein is eliminated while its ribosome-binding site is maintained.
Claim 5
Dependent Claim
27 words
A process according to Claim 4, characterized in that 2 trp regulons are used which are linked in a head to tail fashion according to Fig. 11.
Claim 6
Dependent Claim
32 words
A process according to Claim 2, characterized in that a regulon is used which consists of at least one modified promotor/ribosome-binding site of gene VIII of bacteriophage M13, according to Fig. 10.
Claim 7
Dependent Claim
31 words
A process according to Claim 2, characterized in that a thermosensitive replication mutant is used which is derived from the cloacin DF13 plasmid pVU 208 according to Fig. 12 and 13.
Claim 8
Dependent Claim
67 words
A process according to Claim 2, characterized in that a plasmid is used which is selected from : pUR 1521 (ATCC 39120), 1531, 1541, 1731, 1821, 1831, 1841, 1931 pUR 1522, 1532 , 1542 , 1732 , 1822 , 1832 (ATCC 39197), 1842, 1932 pUR 1523, 1533 (ATCC 39121), 1543, 1733, 1823, 1833, 1843, 1933 pUR 1524, 1534, 1534, 1544, 1734 (ATCC 39198), 1824, 1834, 1844, 1944
Claim 9
Dependent Claim
37 words
A process for producing mammalian chymosin, characterized in that (a) a plasmid according to Claim 2 is incorporated in a micro- organism; (b) the transformed micro-organism is cultivated and (c) chymosin produced by said micro-organism is isolated.
Claim 10
Dependent Claim
16 words
A process according to Claim 9, characterized in that the micro- organisms consist of E-coli cells.
Claim 11
Dependent Claim
27 words
A process according to Claim 9, characterized in that non-toxic edible micro-organisms such as streptococci or lactobacilli, or micro- organisms of bacillus or yeast origin are used.
References Cited (Backward Citations)
No citation records indexed.
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